癌症能够引起人血浆结合珠蛋白（Hp）的糖基化发生异常改变，这种糖基化异常改变可用于筛查高灵敏度和高特异性的肿瘤生物标志物。本项目旨在建立基于亲水作用色谱（HILIC）的糖链高选择性分离方法以及完整糖链结构质谱分析方法的Hp糖基化分析策略，并对癌症状态下Hp糖基化的异常改变进行深入研究。具体来说，首先发展基于新型亲水性固定相的Hp糖链的HILIC分离方法，以实现Hp总糖链以及各糖基化位点上所连接糖链（尤其是糖链异构体）的高效分离；在此基础上发展糖链糖苷键裂解和环内裂解的质谱方法，获取包括单糖组成、连接顺序、分支、连接方式和异头构型在内的Hp糖链完整结构信息，并建立Hp糖链的定量分析方法。最后采用所建立的Hp糖基化分析策略对肝癌患者和慢性肝病患者血清中的Hp糖链差异性进行分析比较，全面剖析癌症状态下Hp所发生的糖基化异常改变，为高灵敏度和高特异性的肝癌生物标志物筛查提供理论依据和研究方法。

The aberrant glycosylation alteration of human plazma protein haptoglobin (Hp) can be utilized for the discovery of novel tumor biomarker with high sensitivity and specificity. The purpose of this project is to establish the Hp glycosylation analysis strategy based on high selective hydrophilic interaction chromatography (HILIC) coupled to full structural mass spectrometry (MS) of Hp glycans,followed by the detailed characterization of the aberrant glycosylation alteration of Hp caused by cancer.First of all,HILIC methods for the separation of Hp entire glycans and glycans attached to each glycosylation site will be developed based on customized hydrophilic stationary phases that have been proved to have high selectivity for the enrichment and separation of oligosaccharides and glycopeptides. Then,qualitative MS methods of glycosidic-bond and cross-ring cleavages of Hp glycans will be developed to aquire the full structural information of Hp glycans, including the monosaccharide composition, sequence, branching, linkage and anomeric configurations. Meanwhile, quantitative MS methods of Hp glycans will be established as well. Finally, the Hp glycosylation analysis strategy will be applied to compare the Hp glycan expression difference between the patients of the hepatocellular carcinoma (HCC) and chronic liver diseases.The project will be helpful to provide the theoretical proof and reseach tools for the screening of HCC biomarkers with high sensitivity and specificity.